Regulation of Microphage Migration Inhibitory Factor (MIF) Activity
a technology activity regulation, which is applied in the field of modulating, neutralizing, or inactivating the activity of migration inhibitory factor (mif), and can solve problems such as unwanted side effects and complications
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example 1
MIF is Only Found Complexed With Other Proteins in vivo
[0041]In order to establish the form of MIF found in tissues and the changes associated with treatments designed to induce neurogenic inflammation; rat intraluminal fluid was analyzed for MIF using ELISA (Table 1). As reported previously, substance P (SP) treatment induced a significant increase in intraluminal MIF amounts (Table 1). Intravesical capsaicin resulted in a dose-dependent excretion of MIF into the intraluminal fluid (Table 1).
TABLE 1Changes in intraluminal MIF concentrationsInflammatory inducerIntraluminal MIF (ng / mL)ControlTreatment(n = 5)(n = 5)Substance P1.8 ± 0.79.5 ± 1.0***(subcutaneous)Control0.1 mM1.0 mM(n = 3)(n = 3)(n = 3)Capsaicin4.1 ± 0.613.9 ± 3.228.6 ± 3.8**(intravesical)Intraluminal MIF amounts were determined using a rat MIF specific ELISA. Data are expressed as mean MIF ng / mL of intraluminal fluid ± S.E.M.,**= p ***= p
[0042]Rat urine, intraluminal fluid and bladder tissue were also analyzed by nativ...
example 2
Identification of MIF Associated Proteins as Alpha-1-Inhibitor-3
[0046]Duplicate samples of intraluminal fluid were separated by non-reducing NuPAGE electrophoresis with one sample analyzed by Western blotting and its duplicate Coomassie stained. X-ray film of the Western blot was used to identify MIF complex Coomassie stained bands for mass spectroscopy analysis (protein Chemistry Core Laboratory; University of Florida, Gainesville, Fla.). These bands were identified as several acute phase proteins (Table 2). The highest and most significant match (for either the 170 or 130 kDa band) was alpha-1-inhibitor-3 (α-1-I-3), however, peptides matching alpha-2-macroglobulin, complement component 4, and ceruloplasmin precursor were also identified.
TABLE 2Proteins identified by mass spectroscopyReferencePeptidesBandNumberProtein (molecular weight)ScoreMatched170 kDagi|12831225α(1)-inhibitor 3 (166 kDa)50120gi|112893α(1)-inhibitor 3 precursor (165 kDa)48717gi|109550α-2-macrog...
example 3
MIF Complexes in Human Urine
Bacterial Cystitis Induced MIF Complexes
[0048]MIF western blot (WB) of urine proteins from UTI patients separated under native conditions resulted in a prominent high molecular weight band in the range of 150 to 500 kDa (FIG. 6). A less intense band at the same molecular weight range was observed in WB of urine from non-UTI patients. Under native WB conditions, a 12 kDa band indicative of monomeric MIF was not detected (FIG. 6). However, under denaturing conditions, WB of the same sample resulted in three prominent high molecular weight bands of 165, 135 and 100 kDa as well as the 12 kDa monomeric MIF band (FIG. 6). MIF staining intensity was greater in the samples from UTI patients. Under reducing conditions only a single MIF staining band at 12 kDa was evident in all the urine samples. The MIF band staining intensity was greatest in urine from UTI patients (FIG. 6).
[0049]Thus, in agreement with Examples 1-2, MIF in human urine is found in high molecular...
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